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A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR
This paper presents a fully automated point-of-care device for protein quantification using short-DNA aptamers, where no manual sample preparation is needed. The device is based on our novel aptamer-based methodology combined with real-time polymerase chain reaction (qPCR), which we employ for very...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9313270/ https://www.ncbi.nlm.nih.gov/pubmed/35884340 http://dx.doi.org/10.3390/bios12070537 |
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author | Cavallo, Francesca Romana Mirza, Khalid Baig de Mateo, Sara Miglietta, Luca Rodriguez-Manzano , Jesus Nikolic, Konstantin Toumazou, Christofer |
author_facet | Cavallo, Francesca Romana Mirza, Khalid Baig de Mateo, Sara Miglietta, Luca Rodriguez-Manzano , Jesus Nikolic, Konstantin Toumazou, Christofer |
author_sort | Cavallo, Francesca Romana |
collection | PubMed |
description | This paper presents a fully automated point-of-care device for protein quantification using short-DNA aptamers, where no manual sample preparation is needed. The device is based on our novel aptamer-based methodology combined with real-time polymerase chain reaction (qPCR), which we employ for very sensitive protein quantification. DNA amplification through qPCR, sensing and real-time data processing are seamlessly integrated into a point-of-care device equipped with a disposable cartridge for automated sample preparation. The system’s modular nature allows for easy assembly, adjustment and expansion towards a variety of biomarkers for applications in disease diagnostics and personalised medicine. Alongside the device description, we also present a new algorithm, which we named PeakFluo, to perform automated and real-time quantification of proteins. PeakFluo achieves better linearity than proprietary software from a commercially available qPCR machine, and it allows for early detection of the amplification signal. Additionally, we propose an alternative way to use the proposed device beyond the quantitative reading, which can provide clinically relevant advice. We demonstrate how a convolutional neural network algorithm trained on qPCR images can classify samples into high/low concentration classes. This method can help classify obese patients from their leptin values to optimise weight loss therapies in clinical settings. |
format | Online Article Text |
id | pubmed-9313270 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-93132702022-07-26 A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR Cavallo, Francesca Romana Mirza, Khalid Baig de Mateo, Sara Miglietta, Luca Rodriguez-Manzano , Jesus Nikolic, Konstantin Toumazou, Christofer Biosensors (Basel) Article This paper presents a fully automated point-of-care device for protein quantification using short-DNA aptamers, where no manual sample preparation is needed. The device is based on our novel aptamer-based methodology combined with real-time polymerase chain reaction (qPCR), which we employ for very sensitive protein quantification. DNA amplification through qPCR, sensing and real-time data processing are seamlessly integrated into a point-of-care device equipped with a disposable cartridge for automated sample preparation. The system’s modular nature allows for easy assembly, adjustment and expansion towards a variety of biomarkers for applications in disease diagnostics and personalised medicine. Alongside the device description, we also present a new algorithm, which we named PeakFluo, to perform automated and real-time quantification of proteins. PeakFluo achieves better linearity than proprietary software from a commercially available qPCR machine, and it allows for early detection of the amplification signal. Additionally, we propose an alternative way to use the proposed device beyond the quantitative reading, which can provide clinically relevant advice. We demonstrate how a convolutional neural network algorithm trained on qPCR images can classify samples into high/low concentration classes. This method can help classify obese patients from their leptin values to optimise weight loss therapies in clinical settings. MDPI 2022-07-19 /pmc/articles/PMC9313270/ /pubmed/35884340 http://dx.doi.org/10.3390/bios12070537 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Cavallo, Francesca Romana Mirza, Khalid Baig de Mateo, Sara Miglietta, Luca Rodriguez-Manzano , Jesus Nikolic, Konstantin Toumazou, Christofer A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR |
title | A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR |
title_full | A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR |
title_fullStr | A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR |
title_full_unstemmed | A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR |
title_short | A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR |
title_sort | point-of-care device for fully automated, fast and sensitive protein quantification via qpcr |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9313270/ https://www.ncbi.nlm.nih.gov/pubmed/35884340 http://dx.doi.org/10.3390/bios12070537 |
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